High-resolution imaging of redox signaling in live cells through an oxidation-sensitive yellow fluorescent protein.
We present the application of a redox-sensitive mutant of the yellow fluorescent protein (rxYFP) to image, with elevated sensitivity and high temporal and spatial resolution, oxidative responses of eukaryotic cells to pathophysiological stimuli. The method presented, based on the ratiometric quantitation of the distribution of fluorescence by confocal microscopy, allows us to draw real-time "redox maps" of adherent cells and to score subtle changes in the intracellular redox state, such as those induced by overexpression of redox-active proteins. This strategy for in vivo imaging of redox signaling circumvents many of the technical limitations currently encountered in the study of complex redox-based phenomena and promises to contribute substantially to this expanding area of signal transduction.
Maulucci
Giuseppe
G
Institute of Physics, Catholic University Medical School, Rome, Italy.
Labate
Valentina
V
Mele
Marina
M
Panieri
Emiliano
E
Arcovito
Giuseppe
G
Galeotti
Tommaso
T
Østergaard
Henrik
H
Winther
Jakob R
JR
De Spirito
Marco
M
Pani
Giovambattista
G
D000818:Animals; D002477:Cells / Q000166:cytology*; D006801:Humans; D008164:Luminescent Proteins; D008722:Methods; D018613:Microscopy, Confocal; D010084:Oxidation-Reduction; D015398:Signal Transduction
D008164:Luminescent Proteins